Abstract

Electrical stimulation of the heel or toes evokes short latency polysynaptic reflexes in muscles of the ankle extensor medial gastrocnemius (MG), the ankle flexor tibialis anterior (TA) and the knee flexor biceps femoris (BF), the co-ordinated actions of which form an organized protective withdrawal response. Previous studies in the rabbit have shown that such reflexes are enhanced (sensitized) or inhibited by application of the chemogenic agent mustard oil (MO) to various areas of the body surface in a manner that reinforces the protective function of these responses. The organization of these ‘sensitization fields’ was strictly controlled by supraspinal pathways from the brain. The aim of the present experiments was therefore to extend these studies of the spatial organization of sensitization of withdrawal reflexes into the rat, the species most commonly used in pain research.

Patterns of facilitation and inhibition of spinal reflexes were obtained and compared in decerebrate spinalized, decerebrate non-spinal, and Alfaxan- anaesthetized rats by applying mustard oil to sixteen different body locations including sites on the ipsilateral and contralateral hindlimbs as well as other off limb areas such as the snout and tail. It was found that in decerebrate spinalized animals, MO application to ipsilateral hindlimb sites enhanced but never inhibited reflex responses in the limb, whilst MO treatment to off limb sites was without effect. In contrast in anaesthetized animals the prevalent effect of MO was inhibition from treatment sites distributed across the entire animal. Reflexes in animals with an intact spinal cord (decerebrate or anaesthetized) were facilitated or inhibited by MO application to ipsilateral hindlimb sites in a way that resembled the modular organization of reflexes per se and previous sensitization studies in the rabbit. However clear differences were also observed in the effects of MO between the two species, including modulation of the heel-MG extensor response in spinalized animals, which in rabbit was inhibited by MO application to the ipsilateral toes whereas in the rat no inhibition by MO was found in spinalized animals. Sensitization of hindlimb reflexes by MO in the rat therefore seems to be influenced by descending inhibitory and facilitatory pathways. These influences were further investigated in subsequent studies.

Whilst the predominant effect of spinalization was a loss of inhibition and an expansion of sensitization fields, in the toes-evoked TA reflex the reverse was noted with regard to MO treatment of distal ipsilateral sites. In this case, facilitation found in non-spinal animals did not occur in the equivalent spinalized cohort, and thereby implies that a descending facilitatory pathway is also implicated in the control of spinal reflex excitability in this model.

In decerebrate rats, the noradrenergic α2-adrenoceptor antagonist RX 821002 or the serotonergic 5-HT3 receptor antagonist ondansetron were administered directly to the spinal cord (intrathecally, i.t.) either alone (dose-response studies) or as a single dose between two successive MO applications to one of three ipsilateral skin sites on the hindlimb (heel, metatarsophalangeal joints or flexion of the ankle). Cumulative i.t. doses of RX 821002 revealed the presence of tonic descending inhibition of all reflex responses as well as preventing MO-evoked inhibition (and possibly facilitation) of reflex responses suggesting the involvement of the α2-adrenoceptor subtype in mediating these effects in this model. On the other hand, cumulative i.t. ondansetron administration resulted in a decrease in the magnitude of reflex responses, thus indicating that 5-HT3 receptors are indeed implicated in tonic descending facilitation of spinal reflexes. In addition i.t. ondansetron revealed that potentiation (and possibly inhibition) of reflexes following an acute chemogenic insult appears to involve the actions of serotonin at 5-HT3 receptors in the spinal cord.

These studies therefore show that the organization of sensitization of hindlimb reflexes in the rat are modulated by supraspinal influences that exist as a balance of descending facilitatory and inhibitory pathways, mediated at least in part by serotonergic 5-HT3 receptors and noradrenergic α2-adrenoceptors.